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Molecular characterization, vector identification and sources of phytoplasmas associated with brinjal little leaf disease in India

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Abstract

Brinjal little leaf (BLL) is a widespread disease of phytoplasma etiology in India that induces severe economic losses. Surveys were conducted in eight brinjal-growing states of India during July 2014 to September 2015 and eighteen BLL samples showing little leaf, phyllody and witches’ broom symptoms were collected for phytoplasma identification. Presence of phytoplasmas was confirmed in all the eighteen BLL samples using polymerase chain reaction with phytoplasma-specific primer pairs (P1/P6, R16F2n/R16R2). Pair wise sequence comparison and phylogenetic relationship of 16S rRNA gene sequences of BLL phytoplasma strains confirmed that sixteen out of eighteen BLL strains belonged to clover proliferation phytoplasma (16SrVI) group and two BLL strains (GKP-A and GKP-B) from Gorakhpur, Uttar Pradesh, were classified under 16SrII group. Further virtual RFLP analysis of 16S rDNA sequences allowed finer classification of BLL strains into 16SrII-D and 16SrVI-D subgroups. BLL phytoplasma strains belonging to 16SrVI-D subgroup were found as the most widespread phytoplasma strains associated with BLL disease in India. 16SrVI-D subgroup phytoplasma association with two symptomatic weed species viz. Cannabis sativa subsp. sativa at Noida, Uttar Pradesh and Portulaca oleracea at IARI fields, New Delhi was also confirmed by nested PCR assays with similar set of phytoplasma-specific primers, pairwise 16S rDNA sequence comparison, phylogeny and virtual RFLP analysis. Out of five identified leafhopper species from BLL-infected fields at Noida, Uttar Pradesh and Delhi, only Hishimonas phycitis was identified as carrier and natural vector of 16SrVI-D subgroup of phytoplasmas by nested PCR assays, sequence comparison, phylogeny, virtual RFLP analysis and transmission assays.

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References

  • Abraham EV, Natarajan K, Murugaesan M (1977) Damage by pests and phyllody to Sesamum indicum in relation to time sowing. Madras Agric J 64:298–301

    Google Scholar 

  • Ahrens U, Seemüller E (1992) Detection of DNA of plant pathogenic mycoplasma like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82:828–832

    Article  CAS  Google Scholar 

  • Anonymous (2013) Agriculture data, Agricultural Statistics databases. http://faostat.fao.org. Organization of the United Nations, Rome, Italy

  • Azadvar M, Baranwal VK (2012) Multilocus sequence analysis of phytoplasma associated with brinjal little leaf disease and its detection in Hishimonas phycitis in India. Phytopathogenic Mollicutes 2:15–21

    Article  Google Scholar 

  • Azadvar M, Baranwal VK, Yadava DK (2009) First report of a 16SrIX (pigeon pea witches’ broom) phytoplasma associated with toria (Brassica rapa cv. toria) phyllody disease in India. New Dis Rep 20:27

    Google Scholar 

  • Bindra OS, Singh B (1969) Biology and bionomics of Hishimonas phycitis (Distant), a jassid vector of little leaf disease of brinjal (Solanum melongena). Indian J Agric Sci 39:912–919

    Google Scholar 

  • Blanche KR, Tran-Nguyen LTT, Gibb KS (2003) Detection, identification and significance of phytoplasmas in grasses in northern Australia. Plant Pathol 52:505–512

    Article  Google Scholar 

  • Chaube S, Kumar S, Dubey D, Tiwari AK, Upadhyaya PP, Rao GP (2015) Identification of a novel phytoplasma (16Sr XIV-A subgroup) associated with little leaf and witches’ broom of Cannabis sativa ssp. sativa and C. sativa ssp. indica in India. Phytoparasitica 43(2):275–279

    Article  CAS  Google Scholar 

  • Deng S, Hiruki C (1991) Amplification of 16S rRNA genes from culturable and nonculturable mollicutes. J Microbiol Methods 14:53–61

    Article  CAS  Google Scholar 

  • Ember I, Munyaneza JE, Crosslin JM, Kölber M (2011) Survey and molecular detection of phytoplasmas associated with potato in Romania and Southern Russia. Eur J Plant Pathol 130:367–377

    Article  Google Scholar 

  • Gopalan C, Rama Sastri BV, Balasubramanian S (2007) Nutritive value of Indian foods. National Institute of Nutrition (NIN), ICMR, Hyderabad

    Google Scholar 

  • Gupta MK, Samad A, Shasany AK, Ajayakumar PV, Alam M (2009) First report of a 16SrVI ‘Candidatus Phytoplasma trifolii’ isolate infecting Norfolk Island pine (Araucaria heterophylla) in India. Plant Pathol 59:399

    Article  Google Scholar 

  • Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucl Acids Symp Ser 4:95–98

    Google Scholar 

  • Kelly PL, Arocha Y, Dider SZ (2009) First report of a 16SrI, ‘Candidatus Phytoplasma asteris’ isolate affecting eggplant and Mikania sp. in Bangladesh. Plant Pathol 58:789

    Article  Google Scholar 

  • Khan MS, Raj SK, Snehi SK (2007) First report of ‘Candidatus Phytoplasma asteris’ affecting sesame cultivation in India. J Plant Pathol 89:301–305

    Google Scholar 

  • Khasa E, Taloh A, Gopala, Prabha T, Rao GP (2016) Molecular characterization of phytoplasmas of ‘Clover proliferation’ group associated with three ornamental plant species in India. 3 Biotech 6:237

    Article  Google Scholar 

  • Kumar J, Gunapati S, Singh SP, Lalit A, Sharma NC, Tuli R (2012) First report of a ‘Candidatus Phytoplasma asteris’ (16SrI group) associated with little leaf disease of Solanum melongena (brinjal) in India. New Dis Rep 26:21

    Article  Google Scholar 

  • Mall S, Upadhyaya PP, Rao GP (2011) Molecular characterization of aster yellows (16SrI) group phytoplasma infecting Cannabis sativa in Eastern Uttar Pradesh. Indian Phytopath 64:132–134

    Google Scholar 

  • Manjunatha N, Prameela HA, Rangaswamy KT, Palanna KB, Wickramaaracgchi WA (2012) Phyllody phytoplasma infecting sesame in South India. Phytopathogenic Mollicutes 2:29–32

    Article  Google Scholar 

  • Maurya R, Madhupriya R, Rao GP (2014) ‘Candidatus Phytoplasma aurantifolia’ related strains affecting two important medicinal plants (Cymbopogon citratus and Tylophora asthmatica) in India. Indian Phytopath 67:303–307

    Google Scholar 

  • Mello AP, Eckstein B, Flores D, Kreyci PF, Bedendo IP (2011) Identification by computer-simulated RFLP of` phytoplasmas associated with eggplant giant calyx representative of two subgroups, a lineage of 16SrIII-J and the new subgroup 16SrIII-U. Int J Syst Evol Microbiol 61:1454–1461

    Article  Google Scholar 

  • Mitra DK (1993) Little leaf, a serious disease of eggplant (Solanum melongena). In: Raychaudhuri SP, Teakle DS (eds) Management of plant diseases caused by fastidious prokaryotes. Associated Publishing Co, New Delhi, pp 73–78

    Google Scholar 

  • Okuda S, Prince JP, Davis RE, Dally EL, Lee IM, Mogen B, Kato S (1997) Two groups of phytoplasma from Japan distinguished on the basis of amplification and restriction analysis of 16S rDNA. Plant Dis 81:301–305

    Article  CAS  Google Scholar 

  • Omar AF, Foissac X (2012) Occurrence and incidence of phytoplasmas of the 16SrII-D subgroup on solanaceous and cucurbit crops in Egypt. Eur J Plant Pathol 133:353–360

    Article  CAS  Google Scholar 

  • Pasquini G, Ferretti L, Barba M (2007) Diffusione del legnonero della vite nel Lazio e caratterizzazione molecolare dell’agente eziologico. Informatore Fitopatologico 4:42–47

    Google Scholar 

  • Priya M, Chaturvedi Y, Rao GP, Raj SK (2010) First report of phytoplasma ‘Candidatus Phytoplasma trifolii’ (16SrVI) group associated with leaf yellows of Calotropis gigantea in India. New Dis Rep 22:29

    Article  Google Scholar 

  • Raj SK, Snehi SK, Kumar S, Banerji BK, Dwivedi AK, Roy RK, Goel AK (2009) First report of ‘Candidatus phytoplasma asteris’ (16SrI group) associated with colour-breaking and malformation of floral spikes of gladiolus in India. Plant Pathol 19:23

    Google Scholar 

  • Rao GP, Mall S, Raj SK, Snehi SK (2010) Phytoplasma diseases affecting various plant species in India. Acta Phytopathol Entomol Hung 46:59–99

    Article  Google Scholar 

  • Rao GP, Madhupriya, Kumar A, Baranwal VK (2015) Classification of the sesame phytoplasma strains in India at the 16Sr subgroup level. J Plant Pathol 97:523–528

    Google Scholar 

  • Salehi M, Izadpanah K, Siampour M, Bagheri A (2007) Transmission of ‘Candidatus Phytoplasma aurantifolia’ to Bakraee (Citrus reticulata hybrid) by Feral Hishimonas phycitis leafhoppers in Iran. Pl Dis 91:466

    Article  Google Scholar 

  • Samad A, Shasany AK, Gupta S, Ajayakumar PV, Darokar MP, Khanuja SPS (2006) First report of a 16SrVI group phytoplasma associated with witches’ broom disease on Withania somnifera. Plant Dis 90:248

    Article  Google Scholar 

  • Samad A, Ajayakumar PV, Shasany AK, Gupta MK, Alam M, Rastogi S (2008) Occurrence of a clover proliferation (16SrVI) group phytoplasma associated with little leaf disease of Portulaca grandiflora in India. Plant Dis 92:832

    Article  Google Scholar 

  • Sertkaya G, Martini M, Musetti R, Osler R (2007) Detection and molecular characterization of phytoplasmas infecting sesame and solanaceous crops in Turkey. Bull Insectol 60:141–142

    Google Scholar 

  • Shantha PR, Lakshmanan M (1984) Vector-borne MLOs of brinjal little leaf. Curr Sci 53:265–267

    Google Scholar 

  • Shukla V, Naik LB (1993) Agro-techniques of solanaceous vegetables. In: Chadha KL, Kalloo G (eds) Advances in Horticulture. Malhotra Pub. House, New Delhi, p 365

    Google Scholar 

  • Siampour M, Izadpanah K, Afsharifar AR, Salehi M, Taghizadeh M (2006) Detection of phytoplasma in insects collected in witches’ broom affected lime groves. Iran J Plant Pathol 42:139–158

    Google Scholar 

  • Singh N, Rao GP, Madhupriya, Upadhayaya PP (2012) ‘Candidatus Phytoplasma trifolii’ associated with little leaf and witches’ broom disease of Datura stramonium L. in India. Phytopathogenic Mollicutes 2:69–71

    Article  CAS  Google Scholar 

  • Smart CD, Schneider B, Blomquist CL, Guerra LJ, Harrison NA, Ahrens U, Lorenz KH, Seemuller E, Kirkpatrick BC (1996) Phytoplasma-specific PCR primers based on sequences of the 16S-23S rRNA spacer region. Appl Environ Microbiol 62:2988–2993

    CAS  Google Scholar 

  • Tamura K, Dudley J, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis (MEGA) software version 5.0. Mol Biol Evol 24:1596–1599

    Article  Google Scholar 

  • Thomas KM, Krishnaswami CS (1939) Little leaf—a transmissible disease of brinjal. Proc Ind Nat Sci Acad 10:201–212

    Google Scholar 

  • Tohidi Z, Salehi M, Ghasemi S, Khanchezar A, Shahamiri SM (2015) Association of a 16SrIX-C phytoplasma with eggplant phyllody in Iran. J Crop Prot 4:247–256

    Google Scholar 

  • Un Nabi S, Dubey D, Madhupriya DD, Rao GP (2015a) Identification of Canabis sativa L. ssp. sativa as putative alternate host of sesame phyllody phytoplasma belongs to 16SrI group in India. Med Plants 7:68–70

    Google Scholar 

  • Un Nabi S, Dubey DK, Madhupriya, Rao GP, Baranwal VK, Sharma P (2015b) Molecular characterization of ‘Candidatus Phytoplasma asteris’ subgroup I-B associated with sesame phyllody disease and identification of its natural vector and weed reservoir in India. Australas Plant Pathol 44:289–297

    Article  CAS  Google Scholar 

  • Varma A, Chenulu VV, Raychaudhuri SP, Prakash N, Rao PS (1969) Mycoplasma like bodies in tissues infected with sandal spike and brinjal little leaf. Indian Phytopath 22:289–291

    Google Scholar 

  • Varma A, Raychaudhuri SP, Chenulu VV, Singh S, Ghosh SK, Prakas N (1975) Proc Indian Nat Sci Acad 41:355–361

    Google Scholar 

  • Verma RK, Dubey GS (1978) Influence of treatment method, duration and concentration of tetracycline on uptake by little leaf affected brinjal plants. Veg Sci 5:36–39

    Google Scholar 

  • Zaim M, Samad A (1995) Association of phytoplasmas with a witches-broom disease of Withania somnifera (L.) Dunal in India. Plant Sci 109:225–229

    Article  CAS  Google Scholar 

  • Zhao Y, Wei W, Lee IM, Shao J, Suo X, Davis RE (2009) Construction of an interactive online phytoplasma classification tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII). Int J Syst Evol Microbiol 59:2582–2593

    Article  CAS  Google Scholar 

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Acknowledgements

The authors are thankful to the Indian Council of Agricultural Research, New Delhi, India, for providing financial assistance during the course of the study. The authors wish to express sincere thanks to the Head, Division of Plant Pathology and the Director, Indian Agricultural Research Institute for providing laboratory facilities. The authors also wish to thank Dr. C.A. Viraktamath, Principal Investigator, Network Project on Insect Biosystematics, Department of Entomology, IARI New Delhi, GKVK, Bangalore, India, for identifying the taxonomy of leafhopper species.

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  1. Plant Virology Unit, Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi, 110 012, India

    Manish Kumar,  Madhupriya & Govind Pratap Rao

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  1. Manish Kumar
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Correspondence to Govind Pratap Rao.

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Kumar, M., Madhupriya & Rao, G.P. Molecular characterization, vector identification and sources of phytoplasmas associated with brinjal little leaf disease in India. 3 Biotech 7, 7 (2017). https://doi.org/10.1007/s13205-017-0616-x

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